Identification of

Medically Important
Fungi

Dr.Kedar Karki
Introduction
 Separation of taxa is still primarily
based on the method of spore
production of the perfect or sexual
state (teleomorph) and three major
subdivisions, the Zygomycetes,
Ascomycetes and Basidiomycetes
are generally recognized. 
Introduction
 However for medical mycologists
two other groups are of particular
importance; the yeasts and the
hyphomycetes or conidial moulds. 
the following groups:
 Dermatophytes
 Yeasts
 Dimorphic Pathogens
 Hyphomycetes (hyaline moulds)
 Hyphomycetes
(dematiaceous moulds)
 Coelomycetes
 Zygomycetes
 Oomycota
  
Dermatophytes

 Microscopic morphology of the

micro and/or macroconidia is the
most reliable identification
character, but you need a good
slide preparation and you may
need to stimulate sporulation in
some strains. 
Dermatophytes

 Culture characteristics such as

surface texture, topography and
pigmentation are variable and are
therefore the least reliable criteria
for identification. 
Dermatophytes

 Clinical information such as the

site, appearance of the lesion,
geographic location, travel history,
animal contacts and race is also
important, especially in identifying
rare non-sporulation species like
M. audouini, T. concentricum and T
schoenleinii etc.  Note: mating
experiments are not practical for
the clinical mycology laboratory.
Three genera are
recognised:

 Epidermophyton:
 Smooth thin-walled Macroconidia
only present, no microconidia,
colonies a green-brown to khaki
colour.
Epidermophyton
floccosum
 On Sabouraud's dextrose agar colonies are
usually slow growing, greenish-brown or khaki
coloured with a suede-like surface, raised and
folded in the centre, with a flat periphery and
submerged fringe of growth. Older cultures
may develop white pleomorphic tufts of
mycelium. A deep yellowish-brown reverse
pigment is usually present. Microscopic
morphology shows characteristic smooth, thin-
walled macroconidia which are often produced
in clusters growing directly from the hyphae.
Numerous chlamydoconidia are formed in
older cultures. No microconidia are formed.
 Culture of Epidermophyton
floccosum.

Macroconidia of E. floccosum

Chlamydoconidia of E. floccosum.
Clinical significance:

 Epidermophyton floccosum is an
anthropophilic dermatophyte with
a world-wide distribution which
often causes tinea pedis, tinea
cruris, tinea corporis and
onychomycosis. It is not known to
invade hair in vivo and no specific
growth requirements have been
reported.
Clinical significance:

 E. floccosum infections may

become epidemic among
personnel using common shower
or gym facilities, e.g. athletic
teams, troops, ship crews and
inmates of institutions.
Microsporum:

 Macroconidia with rough walls

present, microconidia may also be
present.
sp.Teleomorph:
Arthroderma sp.

 Microsporum species form both macro- and

microconidia on short conidiophores.
Macroconidia are hyaline, multiseptate,
variable in form, fusiform, spindle-shaped to
obovate, ranging from 7 to 20 by 30 to 160 um
in size, with thin- or thick- echinulate to
verrucose cell walls. Their shape, size and cell
wall features are important characteristics for
species identification. Microconidia are
hyaline, single-celled, pyriform to clavate,
smooth-walled, 2.5 to 3.5 by 4 to 7 um in size
and are not diagnostic for any one species.
sp.Teleomorph:
Arthroderma sp.

 The separation of this genus from

Trichophyton is essentially based
on the roughness of the
macroconidial cell wall, although in
practice this may sometimes be
difficult to observe. Seventeen
species of Microsporum have been
described, however only the more
common species are included in
these descriptions.
Microsporum spp
 Microsporum audouinii
 Microsporum canis
 Microsorum canis var.
distortum
 Microsporum cookei
 Microsporum equinum
 Microsporum ferrugineum
Teleomorph: Arthroderma
otae

 On Sabouraud's dextrose agar,

colonies are flat, spreading, white
to cream-coloured, with a dense
cottony surface which may show
some radial grooves. Colonies
usually have a bright golden yellow
to brownish yellow reverse
pigment, but non-pigmented
strains may also occur.
 Culture of Microsporum
canis

Macroconidia of M. canis

M. canis growing on boiled polished rice grains

Macroconidia
 Macroconidia are typically spindle-
shaped with 5-15 cells, verrucose,
thick-walled and often have a
terminal knob. A few pyriform to
clavate microconidia are also
present.
Macroconidia
 Macroconidia and/or microconidia
are often not produced on primary
isolation media and it is
recommended that sub-cultures be
made onto boiled polished rice
grains to stimulate sporulation.
Clinical significance:

 Microsporum canis is a zoophilic

dermatophyte of world-wide distribution
which is a frequent cause of ringworm
in humans, especially children. Invades
hair, skin and rarely nails.
 Cats and dogs are the main sources of
infection. Invaded hairs show an
ectothrix infection and fluoresce a
bright greenish-yellow under Wood's
ultra-violet light.
Microsporum spp
 Microsporum fulvum
 Microsporum gallinae
 Microsporum gypseum
 Microsporum nanum
 Microsporum persicolor
Clinical significance:

 The genus Microsporum contains a

number of important species that
are the principle causative agents
of animal and human
dermatophytoses [tinea and
ringworm].
Trichophyton:

 Microconidia present, smooth-

walled macroconidia may or may
not be present.
sp.Teleomorph:
Arthroderma sp.

 The genus Trichophyton is

characterized by the development
of both smooth-walled macro- and
microconidia.
 Macroconidia are mostly borne
laterally directly on the hyphae or
on short pedicels, and are thin- or
thick-walled, clavate to fusiform,
and range from 4 to 8 by 8 to 50
um in size.
 Macroconidia are few or absent in
many species. Microconidia are
spherical, pyriform to clavate or of
irregular shape and range from 2
to 3 by 2 to 4 um in size.
Microconidia, macroconidia
and nodular organs of T.
equinum
Horse infection and
culture of Trichophyton
equinum.
Dimorphic Fungal
Pathogens

 These are fungal infections of the

body caused by fungal pathogens
which can overcome the
physiological and cellular defences
of the normal human host by
changing their morphological
form.  They are geographically
restricted and the primary site of
infection is usually pulmonary,
following the inhalation of conidia. 
Descriptions for the
following fungi are
provided.

 Blastomyces dermatitidis
 Coccidioides immitis
 Histoplasma capsulatum
 Paracoccidioides brasiliensis
Coccidioides immitis

 On Sabouraud's dextrose agar at 25C, colonies

are initially moist and glabrous, but rapidly
become suede-like to downy, greyish white in
color with a tan to brown reverse. However,
considerable variation in both growth rate and
culture morphology has been noted (Rippon,
1988). Microscopic morphology shows typical
single-celled, hyaline, rectangular to barrel-
shaped, alternate arthroconidia, 2.5-4.0 x 3.0-
6.0 um in size, separated from each other by a
disjunctor cell. This arthroconidial state has
been classified in the genus Malbranchea and
is similar to that produced by many non-
pathogenic soil fungi such as Gymnoascus
species.
Culture of Coccidioides
immitis.

Arthroconidia of C. immitis.
Culture of Coccidioides
immitis.
 In the past microscopic
morphology, conversion from the
mould form to the yeast or
spherule form, and animal
pathogenicity have all been used
for identification; however
exoantigen tests are now the
method of choice, at least for
Blastomyces dermatitidis,
Coccidioides immitis and
Histoplasma capsulatum. 
Hyaline Hyphomycetes

 Hyaline Hyphomycetes include

those conidial fungi which are not
darkly pigmented, colonies may be
colorless or brightly colored. 
These include the agents of
hyalohyphomycosis, Aspergillosis,
dermatophytosis and the
dimorphic pathogens, like
Histoplasma capsulatum. 
Descriptions of the following
genera are avaiable:
genera
 AcremoniumAspergillusBeauveria
ChrysosporiumFusariumGeotrichum
GliocladiumGraphiumMadurella
genera
 MalbrancheaPaecilomyces
PenicilliumScedosporium
ScopulariopsisSepedonium
TrichodermaTrichothecium
Verticillium
Identification:

 Identification of the hyphomycetes

is primarily based on microscopic
morphology including; (a) conidial
morphology, especially septation,
shape, size, color and cell wall
texture;
Identification:

 (b) the arrangement of conidia as

they are borne on the
conidiogenous cells, for example
are they solitary, arthrocatenate,
blastocatenate, basocatenate or
gloiosporae etc.,
Identification:

 (c) the type conidiogenous cell, for

example non-specialized or hypha-
like, phialide, annellide or
sympodial etc., and (d) other
additional features such as the
presence of sporodochia or
synnemata.
Culture characteristics,
 although less reliable may also be
useful.  These include surface
texture, topography and
pigmentation, reverse
pigmentation and growth at 37C. 
For identification, potato dextrose
agar and cornmeal agar are two of
the most suitable media to use and
exposure to daylight is
recommended to maximize culture
color characteristics.
  Culture
Characteristics. 
 . Least reliable as the media and growth
conditions play an important part. 
Examine the following characteristics:
 Surface texture [glabrous, suede-like,
powdery, granular, fluffy, downy,
cottony etc].
 Surface topography [flat, raised,
heaped, folded, domed, radial grooved].
  Culture
Characteristics. 
 Surface pigmentation [white,
cream, yellow, brown, green, grey,
black etc].
 Reverse pigmentation [none,
yellow, brown, red etc].
 Growth rate [eg colonies growing
less than 5 mm in 14 days etc].
 Growth temperature studies are
also often very useful [37C, 40C &
45C].
Penicillium sp.
Penicillium sp.

 Colonies are usually fast growing, in shades of green,

sometimes white, mostly consisting of a dense felt of
conidiophores. Microscopically, chains of single-celled
conidia (ameroconidia) are produced in basipetal
succession from a specialized conidiogenous cell called
a phialide. The term basocatenate is often used to
describe such chains of conidia where the youngest
conidium is at the basal or proximal end of the chain. In
Penicillium, phialides may be produced singly, in groups
or from branched metulae, giving a brush-like
appearance known as a penicillus. The penicillus may
contain both branches and metulae (penultimate
branches which bear a whorl of phialides). All cells
between the metulae and the stipes of the
conidiophores are referred to as branches.
Penicillium sp.

 The branching pattern may be either simple

(non-branched or monoverticillate), one-stage
branched (biverticillate-symmetrical), two-
stage branched (biverticillate-asymmetrical) or
three- to more-staged branched.
Conidiophores are hyaline and may be
smooth- or rough-walled. Phialides are usually
flask-shaped, consisting of a cylindrical basal
part and a distinct neck, or lanceolate (with a
narrow basal part tapering to a somewhat
pointed apex). Conidia are globose, ellipsoidal,
cylindrical or fusiform, hyaline or greenish,
smooth- or rough- walled. Sclerotia may be
produced by some species.
Conidiophores of P. verrucosum var. cyclopium
showingtwo-stage branching. Simple conidiophore of P.
cheresanum showing long chains of single-celled

phialoconidia .
Morphological structures and types of conidiophore
branchingin Penicillium. a. simple; b. one-stage branched;

c. two-stage branched; d. three-stage branched

Clinical significance:

 Many species of Penicillium are

common contaminants on various
substrates and are known as
potential mycotoxin producers.
Fusarium sp.

Culture of Fusarium sp.

Culture of Fusarium sp.

 Colonies are usually fast growing,

pale or brightly colored (depending
on the species) and may or may
not have a cottony aerial
mycelium. The color of the thallus
varies from whitish to yellow,
brownish, pink, reddish or lilac
shades. Species of Fusarium
typically produce both macro- and
microconidia from slender
phialides.
 Macroconidia are hyaline, two- to
several-celled, fusiform- to sickle-
shaped, mostly with an elongated
apical cell and pedicellate basal
cell. Microconidia are 1- to 2-
celled, hyaline, pyriform, fusiform
to ovoid, straight or curved.
Chlamydoconidia may be present
or absent.
Microconidia of Fusarium solani are usually
abundant, cylindrical to oval, 1- to 2-celled and
formed from long lateral phialides.
 Macroconidia Fusarium solani are formed after
4-7 days from short multi-branched
conidiophores which may form sporodochia
Chlamydoconidia Fusarium solani are hyaline,
globose,smooth- to rough-walled, borne singly or
in pairs on short lateral hyphal branches, or they
are intercalary.
Clinical manifestations
linical manifestations
 Clinical manifestations of
hyalohyphomycosis caused by
Fusarium; include cutaneous and
subcutaneous infections,
endophthalmitis, osteomyelitis,
and arthritis following traumatic
implantation. Peritonitis has also
been reported in patients on
continuous ambulatory peritoneal
dialysis (CAPD).
Clinical manifestations
 Disseminated infections are similar
to disseminated aspergillosis,
however fungemia and ulcerated
skin lesions are often more
pronounced. The typical patient is
granulocytopenic and receiving
broad-spectrum antibiotics for
unexplained fever.
Penicillium sp. colonizing an orange showing typical green
surface pigmentation with a suede-like surface consisting

of a dense felt of conidiophores.

Culture of Penicillium sp.
on Sabouraud’s dextrose
agar.
Penicillium sp.

 Penicillium sp. showing chains of single

celled phialoconidia (ameroconidia)
produced in basipetal succession from a
specialized conidiogenous cell called a
phialide.  The term basocatenate is
often used to described such chains of
conidia where the youngest conidium is
at the basal or proximal end of the
chain.  In Penicillium phialides may be
produced singly or in groups or from
branched metulae, giving a brush-like
appearance, i.e. a penicillus
Penicillium sp.
Aspergillus fumigatus
Aspergillus fumigatus

 On Czapek dox agar showing

typical blue-green surface
pigmentation with a suede-like
surface consisting of a dense felt
of conidiophores.
Aspergillus fumigatus
Aspergillus fumigatus

 Microscopic morphology of Aspergillus

fumigatus showing typical columnar,
uniseriate conidial heads. 
Conidiophores are short, smooth-walled
and have conical shaped terminal
vesicles, which support a single row of
phialides on the upper two thirds of the
vesicle.  Conidia are produced in
basipetal succession forming long
chains   Conidia are globose to
subglobose, green and rough-walled to
echinulate
Aspergillus fumigatus
Aspergillus niger
Aspergillus flavus
dox agar.  Colonies are
granular, flat, often with
radial grooves, yellow
Mucor sp.
Mucor sp.
 Microscopic morphology of a Mucor
sp. showing erect, simple
sporangiophores forming a
terminal, globose sporangium,
packed with sporangiospores and
with a well developed subtending
columella visible. 
Culture of Candida
albicans

Bubbing yeast cells of Candida albicans